Focused ultrasound stimulation (FUS) is rapidly gaining attention as a non-invasive and highly precise neuromodulatory technique with broad therapeutic potential in neurological and psychiatric disorders. While most reviews to date have emphasized in vivo and clinical studies, the cellular mechanisms underlying FUS remain underexplored. This study presents an innovative and thorough synthesis of FUS effects in in vitro neurological cell models, including SH-SY5Y, PC12, BV2 microglia, NSC-34 motor neurons, and human iPSC-derived neurons and astrocytes. These models offer essential insights into the mechanisms by which FUS influences intracellular calcium dynamics, mitigates oxidative stress, modulates inflammatory responses, and stimulates autophagy, thus facilitating neuroprotection and synaptic resilience in various disease contexts, including Parkinson's disease, Alzheimer's disease, schizophrenia, epilepsy, multiple sclerosis, OCD, and traumatic brain injury. Mapping disease-specific results with comprehensive FUS sonication parameters, this evaluation only focuses on cell-based systems, which is a fundamental advance. Additionally, it emphasizes the incorporation of new technology into FUS, such as acoustically responsive biomaterials, microbubble-assisted gene transfection, and nanoparticle-mediated medication delivery. The study highlights the increasing importance of AI in directing real-time FUS targeting and optimizing parameters, which is leading to tailored neuromodulation treatments. This study establishes a solid groundwork for the advancement of FUS in preclinical research by connecting the dots between cellular bioeffects and translational potential. It highlights the critical need for multidisciplinary methods, standardization, and the use of 3D organoid systems for next-generation brain treatments that fully use FUS.

Department of Animal Physiology Faculty of Science Charles University Albertov 6 Prague 128 00 Czech Republic

Department of Physiology and Biochemistry Faculty of Bioscience Shaheed Benazir Bhutto University of Veterinary and Animal Sciences Sakrand 67210 Pakistan

Division of Experimental Neurobiology Preclinical Research Program National Institute of Mental Health Topolová 748 Klecany 250 67 Czech Republic

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